Assembly of the Bacteriophage T4 Primosome

描述（由申请人提供）： 噬菌体T4复制系统是原核生物和真核生物所采用的DNA复制相关机制中研究得最好的系统之一。这个建议的重点是在DNA复制的情况下，装配和功能的primosome。要完全理解，就需要把原始体的单个蛋白质当作一台大型酶机器来研究。的装配过程中的primosome将询问合奏和单分子FRET平衡和前稳态研究。通过电子显微镜将获得关于原始体的额外结构信息。通过交联/足迹法研究和通过在主动易位和引发期间使用FRET监测蛋白质-蛋白质相互作用来阐明引发体的功能。这个项目的完成将导致详细描述的装配和功能的原始体类似于已经确定的聚合酶全酶。由于DNA复制过程对人类疾病（包括癌症）具有深远的影响，因此了解DNA复制的保真度具有重大的人道主义和经济利益。

DESCRIPTION (provided by applicant): The bacteriophage T4 replication system is one of the best-studied of the related mechanisms of DNA replication employed by prokaryotes and eukaryotes. This proposal is focused on the assembly and functioning of the primosome in the context of DNA replication. A complete understanding will require studying the individual proteins of the primosome as a large enzyme machine. The assembly process of the primosome will be interrogated by ensemble and single-molecule FRET equilibrium and pre-steady state studies. Additional structural information on the primosome will be gained by electron microscopy. The functioning of the primosome will by elucidated by cross-linking/footprinting studies and by monitoring protein-protein interactions using FRET during active translocation and priming by the primosome. Completion of this project will result in a description of the assembly and functioning of the primosome in detail similar to that already determined for the polymerase holoenzyme. Since the process of DNA replication has profound implications for human disease, including cancer, understanding the fidelity of DNA replication is of significant humanitarian and economic interest.